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©SRDE Group, All Rights Reserved. Int. J. Res. Dev. Pharm. L. Sci. 371

International Journal of Research and Development in Pharmacy and Life Sciences Available online at http//www.ijrdpl.com

February - March, 2013, Vol. 2, No.2, pp 371-376 ISSN: 2278-0238

Research Article

ANALYTICAL PROFILE OF RAW MATERIAL AND FINISHED PRODUCT OF CEFUROXIME AXETIL R. Suthakaran, Gopinath*, S.Srikanth, S.B.Muthu Vadivel, Divyatej Nannam

Department of Pharmaceutical Chemistry, Teegala Ram Reddy College of Pharmacy, Saroor nagar, Meerpet, Hyderabad – 97.

*Corresponding Author: Email: [email protected]

(Received: November 05, 2012; Accepted: January 03, 2013)

ABSTRACT

Cefuroxime Axetil is the 1-acetoxyethyl ester of Cefuroxime. Chemically cefuroxime axetil is (RS)-1-hydroxyethyl (6R, 7R)-7-[2-(2-furyl) glyoxyl-amido]-3-(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo-Oct-2-ene-2-carboxylate, 72-(Z)-(O-me-thyl-oxime), 1-acetate-3-carbamate. Its molecular formula is C20H22N4O10S, and it has a molecular weight of 510.48. The cefuroxime raw material was identified by HPLC and IR Spectroscopy. The sample is tested for the solubility, diastereoisomer ratio, crystallinity and bulk density. The amount of cefuroxime axetil was estimated by HPLC assay method. The Cefuroxime axetil finished product was identified by HPLC and IR Spectroscopy. The average weight of tablets was calculated by taking the weights of 20 tablets. The content uniformity of the dosage units was calculated by weight variation method. The dissolution rate was calculated by HPLC method by using Paddle apparatus. The amount of Cefuroxime present in the sample was estimated by HPLC assay method. The known and unknown related substances present in the compound were estimated by HPLC method. Results obtained during the study were satisfactory and can be used for commercial purpose. Keywords: Cefuroxime axetil, HPLC method, IR Spectroscopy.

INTRODUCTION

Cefuroxime1 have a broader spectrum of activity, with

enhanced activity against H.influenzae and the

Enterobacteriaceae resulting from an increased stability to

β-lactamases. Oral agent in this group is the Axetil ester of

Cefuroxime. After oral administration, cefuroxime axetil is

absorbed from the gastrointestinal tract and rapidly

hydrolyzed by nonspecific esterases in the intestinal mucosa

and blood to cefuroxime. During absorption, this acid-stable,

lipophilic, oral prodrug derivative of Cefuroxime Axetil is

hydrolysed to Cefuroxime by intestinal and/or plasma

enzymes. Cefuroxime is subsequently distributed throughout

the extracellular fluids. The axetil moiety is metabolized to

acetaldehyde and acetic acid. The Axetil ester provides an

oral bioavailability of 35 to 50% of Cefuroxime, depending

on conditions. Approximately 50% of serum cefuroxime is

bound to protein. Cefuroxime is excreted unchanged in the

urine. In adults, approximately 50% of the administered

dose is recovered in the urine within 12 hours. In common with

other antibiotics, cefuroxime axetil may affect the gut flora,

leading to lower estrogen reabsorption and reduced

efficacy of combined oral estrogen/progesterone

contraceptives. Adverse reactions to cefuroxime have

Suthakaran et. al., February-March, 2013, 2(2), 371-376


generally been mild and transient in nature.

MATERIALS AND METHODS

1) Cefuroxime axetil raw material

Cefuroxime Axetil Ph. Eur was procured and its molecular

formula C20H22N4O10S and its category Anti-bacterial and

storage conditions preserve in air tight containers protect

from light.

2) HPLC

Instrumentation, Reagents, mobile phase preparation,

chromatographic conditions, preparation of solutions,

evaluation of system suitability and procedure proceed as

directed in the ‘Assay’ The retention time of the principal

peaks for cefuroxime axetil diasteroisomer A and

diasteroisomer B in the chromatogram of the sample solution

should compare to that of sample peaks in the

chromatogram of sample solution.

3) IR

Perkin-Elmer FT-IR, Model- Spectrum One or Equivalent,

Hydraulic pellet press, Agate mortar & pestle, 13mm die

and KBr pellet holder.

3.1) Reagents - Potassium Bromide (KBr) – IR Spectroscopy

grade

3.2) Procedure - Triturate about 2-3mg of sample in 200-

300 mg of previously dried KBr (at 250oC for 1 hour). Make

a pellet and record the IR spectrum between 650 and 4000

cm-1. Do the background correction for KBr using a KBr

pellet. Compare the IR spectrum with similarly recorded

spectrum of Cefuroxime axetil - amorphous working

standard.

4) Diastereoisomer ratio by HPLC

Instrumentation, Reagents, Mobile phase preparation,

Chromatographic conditions, Preparation of solutions,

Evaluation of system suitability and procedure – proceed as

directed in the “Assay”. Calculate the diastereoisomer ratio

from the chromatogram of sample solution as follows:

Diastereoisomer ratio = AT2 /TA

AT2 - Average of peak area counts of the Cefuroxime axetil

diastereoisomer A in the chromatograms of sample solution.

TA - Average to the sum of peak area counts of Cefuroxime

axetil diastereoisomers A and B in the chromatograms of

sample solution.

5) Assay by HPLC

5.1 Reagents

Ammonium dihydrogen orthophosphate, Orthophosphoric

acid, Acetanilide used were analytical grade. Methanol used

was HPLC Grade and water from - Milli-Q.

5.2 Preparation of 0.2M ammonium dihydrogen

phosphate

Dissolve 23.0 gm of ammonium dihydrogen phosphate in

1000mL water. Filter through 0.45µ membrane filter.

5.3 Preparation of mobile phase

Prepare a degassed mixture of 0.2M ammonium dihydrogen

phosphate and methanol in the ratio of 62:38 V/V.

5.4 Preparation of ammonium dihydrogen orthophosphate

buffer

Dissolve 23.0gm of ammonium dihydrogen orthophosphate in

1000mL of water. Adjust the pH to 2.4 ±0.05 with

orthophosphoric acid.

5.5 Chromatographic condition

Column - Zorbax TMS, 5 µ (250x4.6 mm), Pump mode –

Isocratic, Flow rate - 1.5 mL/min, Detection - UV, 278 nm,

Injection volume - 10 µL, Data acquisition time - 25 min.

6) RESULTS AND DISCUSSION

6.1) HPLC Identification

The Retention times of the principal peaks for Cefuroxime

axetil diastereoisomer A and Cefuroxime axetil

diastereoisomer B in the chromatogram of the sample

solution corresponds to that of the sample peaks in the

chromatogram of standard solution.

6.2) IR

The transmission minima or absorption maxima in the

spectrum obtained with the substance recorded as KBr pellet

correspond in position and relative size to those in the

spectrum obtained with the Cefuroxime Axetil Amorphous

Working standard. The amide C=O stretch was found

between 1690-1630 cm-1 in both standard and sample

Cefuroxime axetil. Hence the sample is identified as

Cefuroxime Axetil Raw material. The results were shown in

the IR graph Fig. 1.

6.3) DIASTEROISOMER RATIO BY HPLC

Diastereoisomer ratio = AT2/TA

AT2 - Average of peak area counts of the Cefuroxime axetil

diastereoisomer A in the chromatograms of sample solution.

TA - Average to the sum of peak area counts of Cefuroxime

axetil diastereoisomers A and B in the chromatograms of

sample solution.



AT2 = 1733107 + 1733532= 1733319.5

2

TA = (1802752 + 1733107) + (1803728 + 1733532)/2

= 3536559.5

Diastereoisomer Ratio = 1733319.5/3536559.5 =

049011. The Cefuroxime axetil contains two

diastereoisomers A and B. The diastereoisomer ratio was

calculated by taking the peak areas from HPLC graph as

49011.

6.4) ASSAY BY HPLC

Assay (%m/m, as C20H22N4O10S) (X1) =

(At1×Ws×10×50×50×P)/(As ×50×50×WT×10)

Assay (%m/m, as C20H22N4O10S) (X2) =

(At2×Ws×10×50×50×P)/(As×50×50×WT×10)

At1 - Ratio of Sample Solution-1 (Sum of the peak areas of

the Cefuroxime Axetil Diastereoisomer A and B to the peak

area of Internal Standard) = 4.4655

At2 - Ratio of Sample Solution-2 (Sum of the peak areas of

the Cefuroxime Axetil Diastereoisomer A and B to the peak

area of Internal Standard) = 4.4705

As - Average Ratio of Standard Solution (Sum of the peak

areas of the Cefuroxime Axetil Diastereoisomer A and B to

the peak area of Internal Standard) = 4.43098

Ws - Weight of Standard = 60.86 mg

WT - Weight of Sample = 60.86 mg

P - Potency of Cefuroxime Axetil working standard used =

98.8%

X1 = 4.4655 × 60.86 × 10 × 50 × 50 × 98.8

4.43098 × 50 × 50 × 60.86 × 10

= 99.56%

X2 = 4.4705 × 60.86 × 10 × 50 × 50 × 98.8

4.43098 × 50 × 50 × 60.86 × 10

= 99.68%

Average = X1 + X2 = 99.56 + 99.68 = 99.62%

2 2

Assay (%m/m, as C20H22N4O10S) = 99.62%

The content of Cefuroxime axetil was calculated by HPLC

assay method by using Ammonium dihydrogen

phoasphate:methanol (62:38 v/v). The retention time of

Cefuroxime axetil-A was found at 13.18 min and for

Cefuroxime axetil-B was at 11.34 min. The percentage

potency was calculated by taking the peak areas from the

HPLC graph. The average percentage potency of two

samples was calculated as 99.62%. The results were shown

in Fig. 2, 3, 4 & 5.

7) CEFUROXIME AXETIL FINISHED PRODUCT

IDENTIFICATION

HPLC

The retention time of the principal peaks for Cefuroxime

axetil diastereoisomer A and Cefuroxime axetil

Fig. 1 IR spectra of Cefuroxime Axetil (amorphous)



diastereoisomer B in the chromatogram of the sample

solution corresponds to that of the sample peaks in the

chromatogram of Standard solution.

IR

The transmission minima or absorption maxima in the

spectrum obtained with the substance recorded as KBr pellet

correspond in position and relative size to those in the

Fig. 2: System suitability

Fig. 4: Sample solution

Fig. 3: Standard Cefuroximeaxetil



spectrum obtained with the Cefuroxime Axetil Working

standard. The amine C–N stretch was found between 1340-

1020 cm-1 in both standard and sample Cefuroxime axetil.

Hence the sample is identified as Cefuroxime axetil. The

results were shown in the IR graph Fig. 6.

8) Average Weight

The weights of 20 tablets were mentioned in Table: 1.

Average weight = 997.695 mg. Pass Limit - 1000.0 mg ±

2.0% (980 mg - 1020 mg). The average weight of

Cefuroxime axetil tablets was calculated by taking the

weights of 20 tablets as 997.695 mg. The result obtained

was within the standard limits.

CONCLUSION

In the present work Cefuroxime axetil raw material was

identified by HPLC and IR Spectroscopy. The sample is

tested for the solubility, diastereoisomer ratio, crystallinity,

and bulk density. The amount of cefuroxime axetil was

estimated by HPLC assay method. The Cefuroxime axetil

Finished product was identified by HPLC and IR

Spectroscopy. The average weight of tablets was calculated

Fig. 5: Bracketing Standard

Fig. 6: IR graph of Cefuroxime axetil tablets BP 500 mg



by taking the weights of 20 tablets. The content uniformity of

the dosage units was calculated by weight variation method.

The dissolution rate was calculated by HPLC method by using

Paddle apparatus. The amount of Cefuroxime present in the

sample was estimated by HPLC assay method. The known

and unknown related substances present in the compound

were estimated by HPLC method. In the near future the

process has to be validated for quality assurance of

Cefuroxime axetil tablets.

REFERENCES

1. Graham L. Patric, An Introduction To Medicinal Chemistry, New York, 1995; 154-9,166-88.

2. Burger’s Medicinal Chemistry and Drug Discovery, 6th Edition, 1998; Volume 1: 872-3.

3. Burger’s Medicinal Chemistry and Drug Discovery, 6th Edition, 1998; Volume 5: 639-40.

4. Wilson and Gisvold’s Text Book of Organic Medicinal and Pharmaceutical Chemistry, 11th Edition, 2004; 330.

5. R. M. Silverstein, G. C. Bassler, and T. C. Morrill, Spectrometric Identification of Organic Compounds, 4th Edition, New York, Wiley, 1981; 166.

6. C. P. Sherman Hsu, Handbook of Instrumental Techniques for Analytical Chemistry, 247-70.

7. D.A.Skoog, D.M.West, F.J.Holler, Fundamentals of Analytical Chemistry, Saunders College Publishing.

8. Standard base techniques, High Performance Liquid Chromatography, Drenthe College, The Netherlands.

9. Sandeep Kumar G, Sathish D and Madhusudan Rao Y, “Formulation And Evaluation of Gastroretentive Floating Tablets of Cefuroxime Axetil”, Intl. J. Res. Pharm. Biomed. Sci., Jan – Mar 2012; Vol. 3 (1).

10. Santosh Shelke, Santosh Dongre1, Amit Rathi, Dinesh Dhamecha, Saifee Maria and Mohd Hassan G Dehghan, “Development and Validation of UV Spectrophotometric Method of Cefuroxime Axetil in Bulk and Pharmaceutical Formulation”, Asian J. Res. Chem. 2009; 2(2): 1-11.

Table.8.1. Weights of 20 Tablets

No. Weight of tablet No. Weight of tablet No. Weight of tablet No. Weight of tablet

1 992.50 6 997.54 11 995.67 16 1001.56 2 985.92 7 1001.73 12 999.83 17 997.49 3 1002.73 8 999.74 13 989.58 18 1000.78 4 1001.99 9 1002.84 14 996.99 19 994.55 5 1000.59 10 994.77 15 998.76 20 998.34

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